Nitrogen fixation (acetylene reduction) in the phyllosphere of Thalassia testudinum

N₂ fixation (C₂H₂ reduction) associated with the leaves of the sea grass Thalassia testudinum was investigated at 5 sites in South Florida (Biscayne Bay) and one site in the Bahamas (Bimini Harbor). Significant activities were correlated with the occurrence of a heterocystous blue-green alga (Calothrix sp.) on the leaves. C₂H₂ reduction was not stimulated by organic compounds, either aerobically or anaerobically in the light or dark. Therefore, other physiological types of microbes were not important in N₂ fixation. Diurnal and seasonal variations in N₂ fixation occurred, with maximal rates during the daytime and in the late spring and early summer. N₂ fixation was negligible at four stations in Biscayne Bay. At the fifth station, near Fowey Rock, about 5 kg N ha^-1 year^-1 was fixed. In the summer, the N₂ fixed per day (4–5 mg N m^-2) could provide 4 to 23% of the foliar productivity demands of T. testudinum at this site and the station in Bimini Harbor. N₂ fixation at the periphery of a sea-grass patch, near Fowey Rock, could provide 8 to 38% of the daily nitrogen requirement for leaf production, and thereby might compensate for a less effective trapping and recycling of nitrogen from dead leaves in such regions.     

Grazing effects on nitrogen fixation in coral reef algal turfs

This study addressed whether grazing by the sea urchin Diadema antillarum influenced rates of nitrogen fixation by algal turf communities on Caribbean coral reefs. Because the turfs were nitrogen-limited, we also assessed whether newly-fixed nitrogen was important for supporting net primary productivity by the turfs. We measured acetylene reduction in turfs grown in treatments excluding or including D. antillarum in the presence of other herbivores at 3 m water depth on Tague Bay forereef, St. Croix, U.S. Virgin Islands. These were the first measurements of acetylene reduction on coral reefs under quasi-natural conditions of high water-flow and photosynthetic oxygen generation. Rates of acetylene reduction under these conditions were as high as any measured previously in coral reef communities (mean 7.6 nmol C₂H₄ cm⁻² h⁻¹). Algal turfs grazed by D. antillarum and other herbivores had chlorophyll-specific acetylene reduction rates up to three times higher than when D. antillarum was excluded. High rates of nitrogen fixation by the turfs were sufficient to meet <2% of the nitrogen required to support net chlorophyll-specific primary productivity over 24 h. Grazer-mediated increases in nitrogen fixation do not appear responsible for a parallel enhancement of net primary productivity. Algal turfs at this site must be dependent primarily on external sources of nitrogen. Received: 1 July 1997 / Accepted: 5 September 1997     

Atmospheric dinitrogen fixation by benthic communities of Tikehau Lagoon (Tuamotu Archipelago, French Polynesia) and its contribution to benthic primary production

Acetylene reduction rates were measured in lagoonal sediments, cyanobacterial mats and limestone surfaces between 1991 and 1995 at many sites, depths and seasons; all the studied substrata contained cyanobacteria. The acetylene reduction/¹⁵N₂ fixation ratio was measured for the different communities and varied between 1.8 and 4.8, depending on substratum. Fixation rates were 1.7 to 7 times higher during daylight compared to night-time rates. N₂ fixation rates ranged from 0.4 to 3.9 mg N m^-2 day^-1 for the lagoonal sediment/mat communities, and the rate was about 2 mg N m^-2 day^-1 for the lagoonal limestone substrata. Total lagoonal benthic N₂ fixation contributed 24.4% of the total nitrogen requirement for the benthic primary production of benthic communities of the lagoon. The input of N₂ fixation by the microbial planktonic communities (including cyanobacteria) of the lagoon, which are highly productive, is unquantified but is likely to be large.     

Effects of oxygen,mannitol and ammonium concentrations on nitrogenase [C2H2] activity in a marine skeletal carbonate sand

Following a lag of 3 to 18 h, acetylene reduction in mannitol-amended sand systems proceeded at approximately constant and high rates for periods up to 4 days. Carbon dioxide production and O₂ consumption were low in these systems in comparison to similar systems additionally amended with ammonium, indicating N-limitation of growth in the former. Thus, long-term acetylene assays of mannitol-amended sand and suspensions from the sand incubated at various partial pressures of oxygen could be used to characterize the O₂-sensitivity of the N₂-fixing bacterial population as a whole, in batch-type systems with a minimal degree of enrichment or change in pO₂ during the course of the assays. Results of various studies suggested that aerobic or microaerophilic N₂-fixing bacteria were absent or scarce in the sand, and that nitrogenase activity occurring in aerobically incubated systems occurred in anaerobic microenvironments. Hydrogen stimulated acetylene-reducing activity, but the time course differed from that of mannitol-supported activity, and proceeded with shorter lags in systems incubated at 0.2 and 0.05 atm O₂ than in systems incubated anaerobically. Efficiency of N₂ fixation [C₂H₂] increased with decreasing initial mannitol concentration. For sand washed with seawater to remove native combined inorganic nitrogen, and amended with 0.015% mannitol, 374 μmoles added NH₄-N/kg wet sand caused almost complete repression of nitrogenase activity, while concentrations as low as 12 μmoles added NH₄-N/kg wet sand appeared to cause at least partial repression of nitrogenase activity. Some implications of these results for the existence of anaerobic microenvironments in the cavities of skeletal carbonates, and for N₂-fixation in the seagrass rhizosphere are discussed.     

Use of the “Acetylene blockage” technique for assaying denitrification in a salt marsh

The acetylene blockage technique was evaluated for measurement of denitrification in salt-marsh sediments (near Halifax, Nova Scotia, Canada). N₂O in the gas phase of closed Spartina alterniflora marsh-sediment systems was analyzed with use of a thermal conductivity gas chromatograph sensitive to approximately 0.1 nmoles ml^-1 gas. No N₂O was detected for unfertilized sediment samples taken through the growing season and incubated in sealed buckets with 10% C₂H₂. For sediment samples amended with nitrate and for enrichments, initial rates of N₂O evolution were higher in the presence of 10% C₂H₂ than in the absence of C₂H₂, but after longterm incubation N₂O was consumed in some samples containing C₂H₂ as well as in samples without C₂H₂. In addition, total gaseous nitrogen (N₂ and N₂O) production in the absence of C₂H₂ was higher than in the presence of C₂H₂. Acetylene appears to be an inconsistent inhibitor of N₂O reduction in salt-marsh sediments. The usefulness of the acetylene-denitrification technique in this habitat is, therefore, questionable.     

High rates of nitrogen fixation on coral skeletons after predation by the crown of thorns starfish Acanthaster planci

At One Tree Reef, Great Barrier Reef, Australia, between 1983 and 1985, corals killed by the crown of thorns seastar Acanthaster planci L. gave rise to skeletons which were colonised rapidly by blue-green and other algae. For the next 3 to 9 mo these coral skeletons showed over three times more nitrogen fixation (acetylene reduction) than control substratum rates (9 to 32 nmol vs 3 to 10 nmol C₂H₂ cm^-2 h^-1, over all seasons). These values convert to relatively high annual fixation rates of 37 to 127 kg N ha^-1 yr^-1 but, at the low densities of A. planci on One Tree Reef (ca. 0.65 ha^-1), this has little impact on the total nitrogen fixation rate and, as a result, on the level of organic nitrogen in the system. However, it is suggested that on reefs subjected to high aggregations of a. planci such an effect would enhance the level of organic nitrogen and lead to greater primary and secondary production throughout the reef system.     

Seasonal variations in nitrogen-fixation (acetylene reduction) and sulphate-reduction rates in the rhizosphere of Zostera noltii: nitrogen fixation by sulphate-reducing bacteria

Nitrogen-fixation (acetylene reduction) rates were measured over an annual cycle in meadows of the seagrass Zostera noltii Hornem in the Bassin d'Arcachon, south-west France, between March 1994 and February 1995, using both slurry and whole-core techniques. Measured rates using the slurry technique consistently overestimated those determined on whole cores, probably due to the release of labile organic carbon sources as a result of root damage during preparation of the slurries. Thus, the whole-core technique may provide a more accurate estimate of in situ activity, since disturbance of physicochemical gradients of oxygen, sulphide, nutrients and the relationship between the plant roots and the rhizosphere microflora is minimised. Rates measured by the whole-core method were 1.8- to 4-fold greater (dependent upon season) in the light than those measured during dark incubations, indicating that organic carbon diffusing from the plant roots during photosynthesis was an important factor in regulating nitrogen fixation in the rhizosphere. Additions of sodium molybdate, a specific inhibitor of sulphate-reducing bacteria (SRB) inhibited acetylene-reduction activity by >80% as measured by both the slurry and whole-core techniques throughout the year, inferring that SRB were the dominant component of the nitrogen-fixing microflora. A mutualistic relationship between Z. noltii and nitrogen-fixing SRB in the rhizosphere, based on the exchange of organic carbon and fixed nitrogen is proposed. Acetylene- and sulphate-reduction rates showed distinct summer peaks which correlated with a reduced availability of ammonium in the sediment and the annual growth cycle of Z. noltii in the basin. Overall, these data indicate that acetylene reduction (nitrogen fixation) activity in the rhizosphere of Z. noltii was regulated both by the availability of organic carbon from the plant roots and maintenance of a low NH ₄ ⁺ concentration in the vicinity of the plant roots due to efficient assimilation of NH ₄ ⁺ by Z. noltii during the growth season. Nitrogenfixation rates determined from acetylene-reduction rates measured using the whole-core technique ranged from 0.1 to 7.3 mg N m^-2d^-1, depending on season, and were calculated to contribute between 0.4 and 1.1 g N m^-2yr^-1, or 6.3 to 12% of the annual fixed nitrogen requirement of Z. noltii.     

Hydrodynamics,diffusion-boundary layers and photosynthesis of the seagrasses Thalassia testudinum and Cymodocea nodosa

Photosynthetic rates of aquatic plants frequently increase with increasing current velocities. This is presumably due to a reduction in the thickness of the diffusion boundary-layer which allows for a higher carbon availability on the plant surface. Blades of the seagrasses Thalassia testudinum and Cymodocea nodosa exposed to different current velocities under controlled laboratory conditions, showed increased photosynthetic rates with increasing flow only at low current velocities (expressed as blade friction velocities, u _*). Carbon saturation of photosynthetic processes occurred at a relatively low u _* level (0.25 cm s^-1) for T. testudinum collected from a calm environment compared to C. nodosa (0.64 cm s^-1) collected from a surf zone. No further enhancement of photosynthetic rates was observed at higher u _* levels, suggesting limitations in carbon diffusion through the boundary layer below critical u ^* levels and possible limitations in carbon fixation by the enzymatic system at higher u _* levels. These results, as well as those of previous theoretical studies, assumed the flow on the immediate seagrass-blade surface to be hydrodynamically smooth. The presence of epiphytes and attached debris causes the surface of in situ seagrass blades to be exposed to flows ranging from smooth to rough-turbulent. As a consequence, the boundary-layer thickness on moderately epiphytized blades under medium to high flow-conditions is not continuous, but fluctuates in time and space, enhancing carbon transport. In situ u _* levels measured directly on blades of seagrasses indicate that T. testudinum and C. nodosa can be exposed to conditions under which the boundary layer limits photosynthesis during short periods of time (milliseconds) during low-energy events. As waves cause the thickness of the diffusion boundary-layer to fluctuate constantly, carbon-limiting conditions do not persist for prolonged periods.     

Nitrogen fixation in the North Pacific Ocean

Nitrogen fixation in the euphotic zone of the ocean was measured by C₂H₂ reduction and ¹⁵N₂ incorporation associated with Trichodesmium sp. and also with Richelia intracellularis occurring within the cells of Rhizosolenia styliformis var. longispina, and R. cylindrus. The vertical distribution of N₂ fixation activity, N₂-fixing species, particulate matter and dissolved nutrients was measured. The effects of light intensity, sample concentration, length of incubation, and nutrient enrichment on the rates of C₂H₂ reduction were determined. Estimates of the importance of N₂ fixation in adding previously uncycled nitrogen to the euphotic zone are given.     

Spatial variations in the N2O emissions and denitrification potential of riparian buffer strips in a contaminated urban river

Spatial variations in the N₂O emissions and denitrification potential of riparian buffer strips (RBS) in a polluted river were examined. The river received large pollutant inputs from urban runoff and wastewater discharge, resulting in impaired water quality in the river and downstream reservoir. The potential for nitrogen removal by RBS was evaluated by measuring in situ N₂O emission fluxes in static closed chambers and sediment denitrification potentials with acetylene inhibition techniques. The results showed that N₂O emission fluxes decreased from the upstream (16.39 μg/(m²·h)) to downstream (0.30 μg/(m²·h)) sites and from the water body to upland sites. The trend in decreasing N₂O emission fluxes in the downstream direction was mainly associated with sediment/soil textures (clay loam→sandy soil) and sediment/soil water contents and was also related to the vegetation along the RBS and nutrients in the sediments/soils. The correlation coefficient was highest (r=0.769) between the N₂O emission flux and sediment/soil water content. Sediment/soil denitrification potentials under N-amended and ambient conditions were higher (highest 32.86 mg/(kg·h)) for the upstream sites, which were consistent with in situ N₂O flux rates.     

Nitrogen fixation by blue-green algae associated with the siphonous green seaweed Codium decorticatum: effects on ammonium uptake

Nitrogen fixation (acetylene reduction) at rates of up to 1.2 g N₂ g dry wt^-1 h^-1 was measured for the siphonous green seaweed Codium decorticatum. No nitrogenase activity was detected in C. isthmocladum. The nitrogenase activity was light sensitive and was inhibited by the addition of DCMU and triphenyl tetrazolium chloride. Additions of glucose did not stimulate nitrogen fixation. Blue-green algae (Calothrix sp., Anabaena sp., and Phormidium sp.) were implicated as the organisms responsible for the nitrogenase activity. They occurred in a reduced microzone within the C. decorticatum thallus where nitrogen fixation was optimized. Nitrogen fixation did not affect the kinetic constants for ammonium uptake in C. decorticatum (K_s=12.0 M, V_max=13.4 mol NH₃ g dry wt^-1 h^-1) determined using the perturbation method. Nevertheless, C. decorticatum thalli which fixed nitrogen had internal dissolved nitrogen concentrations which were over 1.4 times higher than in non-fixing thalli. This suggests that if C. decorticatum does derive part of its nitrogen requirement from the blue-green algae which it harbors, the transfer does not involve competition between this process and the uptake of ambient ammonium.     

Nitrogen fixation (acetylene reduction) associated with the living coral Acropora variabilis

To estimate N₂-fixation, acetylene reduction assays were carried out on portions of the branches of the coral Acropora variabilis from the west coast of Malaysia. In some experiments, a sub-surface incubation apparatus was employed that was designed to keep the coral fragments near to their natural depth of occurrence. Other shipboard experiments used metabolic inhibitors to investigate the class of organism reducing acetylene. Stumps of coral gave the highest rates of activity, probably attributable to loosely associated cyanophytes. Coral tips also reduced acetylene at relatively high rates; reduction was enhanced in light by increased CO₂ concentration and decreased O₂ tensions indicative of photosynthetic bacteria. Algal material was not obvious on the tip surfaces and so the active organism was probably more integral to the coral structure than it was in the stumps. Maximum rates of acetylene reduction measured translated to 2.5 mg N₂ fixed per outcrop per day.     

Thalassia testudinum productivity and grazing by green turtles in a highly disturbed seagrass bed

There has been an historical decline in the seagrass beds in Maho and Francis Bays, St. John, U.S. Virgin Islands: presently (1986) there are only five small seagrass beds in shallows water. These seagrass beds are highly disturbed by heavy boat usage and are intensively grazed by the green turtle Chelonia mydas L. Fifteen to 50 boats anchor each night in the bays: anchor scars cause a loss of up to 6.5 m² d^-1 or 1.8% yr^-1 of the seagrass beds. Seagrasses regrew into such scars only minimally within a period of 7 mo. The size of the green turtle population was estimated at 50 subadults and their feeding behavior was determined by direct observation and radiotelemetry. The behavior of the green turtles differed from other observations published on the species. Here, the turtles grazed all available Thalassia testudinum, their preferred seagrass food, rather than creating discrete grazing scars, and spent all their waking hours (9 h per day) feeding. Areal productivity of T. testudinum leaves (33 to 97 mg dry wt m^-2d^-1) in the bays was at least an order of magnitude lower than published values or than the productivity of another, lessdisturbed seagrass bed on St. John, despite having comparable leaf-shoot density. Leaf shoots were stunted, fragile, achlorotic, and had only two leaves as opposed to the five leaves per shoot more typically seen. The green turtle population was near the estimated carrying capacity of T. testudinum, based on the standing crop and productivity of T. testudinum and the grazing rate of the turtles. The effect of disturbance of T. testudinum from boats and turtles was assessed by excluding these with emergent fences. Within 3 mo of protection, the areal and shoot-specific productivity of T. testudinum leaves as well as leaf size increased significantly compared to unprotected areas. Conservation efforts are recommended in Maho Bays and Francis because seagrass productivity is low, disturbance rates are higher than recovery rates, the turtles cannot increase further their feeding rate in order to compensate for such factors, and there are few alternate sources of T. testudinum on the north shore of St. John.Contribution No. 175 from West Indies Laboratory, Teague Bay, Christiansted, St. Croix, U.S. Virgin Islands 00820, USA     

Nitrogen fixation (acetylene reduction) associated with macroalgae in a coral-reef community in the Bahamas

N₂ fixation (C₂H₂ reduction) was associated with several species of macroalgae on a coral reef near Grand Bahama Island. The highest rates were associated with Microdictyon sp. (Chlorophyceae) and Dictyota sp. (Phaeophyceae). Extensive mats of filamentous blue-green algae, not heterotrophic bacteria, were the N₂ fixing agents: in experiments with samples of Microdictyon sp., the activity was lightdependent and not stimulated by organic compounds under either aerobic or anaerobic conditions. Assays in situ, at 20 m depth, and on shipboard, gave similar rates of N₂ fixation; the cyanophytes presumably have pigment adaptations to function in blue light. The maximum rate of N₂ fixation, associated with Microdictyon sp., was 3.8 g N fixed g dry weight^-1 h^-1. Coral-reef communities flourish in nutrientimpoverished waters, and therefore any input of nitrogen is probably important in stabilizing such ecosystems.     

Nitrogen cycling in microbial mats: rates and patterns of denitrification and nitrogen fixation

Spatial and temporal variations in nitrogen fixation and denitrification rates were examined between July 1991 and September 1992 in the intertidal regions of Tomales Bay (California, USA). Microbial mat communities inhabited exposed mudflat and vegetated marsh surface sediments. Mudflat and marsh sediments exhibited comparable rates of nitrogen fixation. Denitrification rates were higher in marsh sediments. Nitrogen fixation rates were lowest during January at both sites, whereas highest rates occurred during summer and fall. Denitrification rates were highest during fall and winter months in marsh sediments, while rates in mudflat sediments were highest during summer and fall. In mudflat sediments, nitrogen fixation and denitrification rates, integrated over 24 h, ranged from 6 to 79 mg N m^-1 d^-1 and 1 to 10 mg N m^-2 d^-1, respectively. Rates of denitrification represented between 6 and 20% of nitrogen fixation rates during the day, but exceeded or were equivalent to nitrogen fixation rates at night. The highest integrated rates of both nitrogen fixation and denitrification occurred during July, whereas, the highest percent loss occurred during spring when denitrification rates amounted to 20% of nitrogen fixation rates during the day. Over an annual cycle, inputs of fixed N to mudflat communities occurred exclusively during daylight. These results underscore the importance of determining integrated diel rates of both nitrogen fixation and denitrification when constructing N budgets. Using this approach, it was shown that microbial denitrification can represent a significant loss of combined nitrogen from mats on daily as well as monthly time scales.     

Aerobic nitrogenase activity measured as acetylene reduction in the marine non-heterocystous cyanobacterium Trichodesmium spp. grown under artificial conditions

Aerobic nitrogenase activity in the marine non-heterocystous cyanobacterium Trichodesmium spp. NIBB 1067, isolated off the Izu Peninsula, Japan in 1983 and grown under artificial conditions, was assayed by the acetylene reduction method. This strain exhibited acetylene reduction activity under aerobic conditions when cells had been grown in the medium free of combined nitrogen. Activity was markedly enhanced by light, and dependent on the growth phase being higher during the exponential growth phase and lower during the late linear and stationary growth phases. Since typical colony formation occurred during the last growth phase, the present results contradict the idea that N₂-fixation depends on colony formation. The photosynthesis inhibitor DCMU at 10^-6 M inhibited light-dependent acetylene reduction completely. Acetylene reduction by Trichodesmium spp. was tolerant of O₂ as strongly as that in the heterocystous cyanobacteria. Even at a partial pressure of oxygen (p_{O 2}) of 3 atm, the activity still remained as high as half of the maximum. It was almost under anaerobic conditions. Maximum activity was obtained at pO₂ of ca. 0.1 atm.     

Acetylene reduction (nitrogen fixation) in a Delaware,USA salt marsh

Acetylene reduction (nitrogen fixation) was measured in several vegetational areas in a Delaware, USA salt marsh. Samples were collected for 1 yr and the results showed a seasonally variable pattern of acetylene reduction at all stations. Peak rates were generally recorded during the later summer and early fall (September–October). The seasonality was influenced mainly, although not exclusively, by the soil temperature. In addition, samples collected in short Spartina alterniflora stands exhibited rates which were up to 20-fold higher than those found in samples from tall S. alterniflora stands. Over 50% of the total yearly ethylene production occurred from mid-August until the beginning of December at the tall and short S. alterniflora stations. Maximum activity occurred at 5 cm depth for all stations. Surface activity accounted for only 3–4% of the total measured in the top 20 cm. Addition of glucose or mannitol resulted in considerable increases in activity, thus suggesting that heterotrophic acetylene reduction is carbon and/or energy limited. The results obtained in this study indicate that the measured rates are only potential rates and that considerable caution must be used in extrapolating from acetylene reduction rates to nitrogen fixation rates in situ.     

Aerobic nitrogen fixation by the marine non-heterocystous cyanobacterium Trichodesmium (Oscillatoria) spp.: Its protective mechanism against oxygen

Nitrogen fixation (acetylene reduction) by the marine non-heterocystous cyanobacteria, Trichodesmium thiebautii and T. erythraeum, is sensitive to oxygen. Its sensitivity to oxygen was intensified when the colonies of T. thiebautii were disintegrated, but the separate trichomes yielded still retained the capacity for light dependent acetylene reduction. Trichodesmium colonies evolved hydrogen under argon in the light. The addition of carbon monoxide with DCMU [3-(3,4-dichlorophenyl)-1,1-dimethylurea] enhanced the rate of hydrogen evolution to approximately the same level as that of the maximum acetylene reduction on an electron basis. This probably results from the inhibition of the uptake hydrogenase. We propose that the uptake hydrogenase functions to protect nitrogenase from damage by oxygen.     

Heterotrophic nitrogen fixation (acetylene reduction) during leaf-litter decomposition of two mangrove species from South Florida, USA

Heterotrophic nitrogen-fixation (acetylene reduction) was measured during decomposition (under dark conditions) of Rhizophora mangle L. and Avicennia germinans (L.) Stearn leaf litter. Nitrogen-fixation rates in leaf litter increased following 24 d incubation, then decreased after ≃44 d for both species. Maximum rates of 66.2 and 64.6 nmol C₂H₄ g⁻¹ dry wt h⁻¹ were reached by R. mangle and A. germinans leaf litter, respectively. Higher fixation rates of leaf litter were associated with an increase in water content and sediment particles on leaf surfaces of both species. Rates of nitrogen fixation by diazotrophs attached to sediment particles were not significantly different from zero. With additions of d-glucose, ethylene production rates increased by factors of 625-, 34- and 7-fold for sediment, R. mangle and A.␣germinans leaf litter, respectively, compared to rates prior to enrichment. These organically enhanced rates of nitrogen fixation on leaves could be accounted for by increased activity associated with attached sediment particles and not the leaf material. Total phenolics [reported as tannic acid equivalent (TAE) units] decreased nitrogen-fixation rates when added to d-glucose-enriched sediment at >20 mg TAE l⁻¹. Phenolic compounds could explain the initial lag in rates of nitrogen fixation during leaf-litter decomposition of R. mangle (initial content of 110.8 mg TAE g⁻¹ dry wt), but not of A. germinans (initial content of 23.4 mg TAE g⁻¹ dry wt). The higher phenolic content and reportedly lower carbon substrate of R. mangle did not result in species-specific differences in either the magnitude or temporal pattern of nitrogen fixation compared to A. germinans leaf litter. We conclude that the availability of organic substrates leached from the leaf litter along with colonization by the heterotrophic diazotrophs (as indicated by sediment accumulation) controls nitrogen-fixation rates in a similar manner in the leaf litter of both species. Received: 8 August 1997 / Accepted: 4 December 1997     

Nitrogen fixation on a coral reef

Acetylene reduction was used to assess nitrogen fixation on all major substrates at all major areas over a period of 1 to 6 yr (1980–1986) at One Tree Reef (southern Great Barrier Reef). Experiments using ¹⁵N₂ gave a ratio of 3.45:1.0 for C₂H₂ reduced:N₂ fixed. Acetylene reduction was largely light-dependent, saturated at 0.15 ml C₂H₂ per ml seawater, and linear over 6 h. High fixation was associated with two emergent cyanophyte associations, Calothrix crustacea and Scytonema hofmannii, of limited distribution. Subtidally, the major contribution to nitrogen fixation came from well-grazed limestone substrates with an epilithic algal community in the reef flat and patch reefs (3 to 15 nmol C₂H₄ cm^-2 h^-1). Similar substrates from the outer reef slope showed lower rates. Nitrogen fixation on beach rock, intertidal coral rubble, reef crest and lagoon sand was relatively small (0.3 to 1.0 nmol C₂H₄ cm^-2 h^-1). Seasonal changes in light-saturated rates were small, with slight reduction only in winter. Rates are also reported for experimental coral blocks (13 to 39 nmol cm^-2 h^-1) and for branching coral inside and outside territories of gardening damselfish (3 to 28 nmol cm^-2 h^-1). This work supports the hypothesis that the high nitrogen fixation on the reef flat and patch reefs of the lagoon (34 to 68 kg N ha^-1 yr^-1) is because these subtidal areas support highly disturbed communities with the greatest abundance of nitrogen-fixing cyanophyte algae. It is calculated from a budget of all areas that One Tree Reef has an annual nitrogen fixation rate of 8 to 16 kg N ha^-1 yr^-1.